• Reproductive and Developmental Biology
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• v.36 no.3
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• pp.147-154
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• 2012

This study was designed to determine whether low-density lipoproteins (LDL) from egg yolk and taurine, hypotaurine and trehalose as antioxidant in extender improve the freezability and fertility of Korean Jeju Black Bull semen. The semen was cryopreserved with tris egg yolk extenders containing 7% glycerol and treated 4% LDL, 20 mM taurine, hypotaurine and trehalose. Frozen-thawed sperm were evaluated motility, viability, membrane, and acrosome integrity and sperm penetration ability. The results were compared to semen cryopreserved in tris egg yolk extender only as control. Frozen-thawed semen evaluation cleary indicated that the addition of LDL and LDL-antioxidants (taurine, hypotaurine and trehalose) combination were significantly improved (p<0.05) the viability (%; with staining test using eosin-Y) compared to control spermatozoa. Also, in membrane integrity (%; with supravital hypo-osmotic swelling test), not only LDL-antioxiants combination but also LDL were significantly increased (p<0.05) the swelled sperm using HOST compared to control. Sperm acrosome integrity state was classified by CTC (chlortetracycline) staining test. F pattern was significantly increased in LDL-antioxidant combination than control (p<0.05) and B pattern was not significantly differences among all treatments and control. However, AR pattern was significantly decreased in LDL-antioxidants combination than control (p<0.05). Pronucleus formation and sperm penetration index (SFI) were significantly increased in LDL and LDL-antioxidants combination than control (p<0.05). Especially, LDL-taurine significantly improved pronucleus fomation and SFI than LDL (p<0.05). It was concluded that LDL and LDL-antioxidants in extender improved the freezability and fertility of Korean Jeju Black bull spermatozoa.

• Journal of Embryo Transfer
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• v.22 no.3
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• pp.173-178
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• 2007

돼지 정자의 성 선별에는 일반적으로 유속 세포 분석기를 이용한다. 유속 세포 분석은 DNA량의 차이에 기초하여 정자를 분리하는 기술로써 X 정자와 Y 정자를 90% 정확도로 분리할 수 있다. 그러나 이러한 유속 세포분석 기술은 정자의 손상을 야기해 정자의 기능과 수정능에 영향을 미치므로, 본 연구에서는 특정한 핵산 서열을 탐지할 수 있는 Chromogenic in situ hybridization(CISH)을 그와 비교하여 평가하였다. 유속 세포 분석을 수행하기 위해 정자를 SYBR 14와 PI로 염색하였고, histogram, dotplot, density, contour를 측정하였다. Y 염색체 특이적인 primer를 이용한 PCR로 유속 세포 분석의 정확도를 검사하였다. HRP/DAB 시스템에 기초한 CISH 분석에는 X 또는 Y 염색체에 상보적으로 결합하는 probe가 사용되었다. CISH 분석은 기존의 방법들보다 빠르고 쉬우며 비용이 적게 든다는 장점이 있다. 또한, CISH는 보다 정화한 정자의 선별을 가능하게 하는 것으로 나타났다. 본 연구에 따르면 CISH가 기존의 선별 방법들을 평가하는 기술로서만이 아니라 특정한 성별을 가진 포유동물의 생산에도 사용될 수 있을 것이다.

• Clinical and Experimental Reproductive Medicine
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• v.18 no.1
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• pp.73-80
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• 1991

We compared fertilizing potential measurements by the zona-free hamster egg penetration assay with the in vitro fertilization and embryo transfer program was evaevulated for their ability to fertilize zona free hamster egg. Spermatozoa from 12 presumeably fertile donors and from the male partners of 56 infertile couples were evaluated for their ability to fertilizing potentials. Penertration rates of fertile donors were $36.2{\pm}27.7%$ ; Fertilization rates of infertile couples between with normal semen parameters and with abnormal semen parameters were $28.7{\pm}19.1$, $5.7{\pm}8.9%$, respectively. Sperm motility of couples with penetration rates between on 15-30% and on 30> were $54.1{\pm}4.6$, $55.5{\pm}8.3%$ respectively. Hamster penetration rates of couples participating in an in vitro fertilization and embryo transfer program was $38.9{\pm}29.9%$. But in one case, a positive fertility assessment was obtained in the absence of fertilization of the wife's eggs attributable to egg immaturity. This method may have potential value as a diagnostic tool in evaluation human sperm fertilization capacity which avoids the ethical and logistical problems associated with fertilizing of human eggs in vitro.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.10
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• pp.1276-1281
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• 2010

The present study was conducted to investigate the effects of butylated hydroxytoluene (BHT) supplementation on diluted, cooled and frozen-thawed ram spermatozoa. After primary evaluation of collected ejaculates, only semen samples with motility of more than 70% and sperm concentration higher than $3{\times}10^3$ sperm/ml were used for cryopreservation. The selected semen samples were then pooled and diluted 1:4 with Tris Citrate Fructose Yolk (TCFY) extender supplemented with different concentrations of BHT (0.5, 10, 2.0 and 3.0 mM). As the control, semen was diluted and frozen in the diluent without BHT. Motility, progressive motility, viability, membranes and acrosome integrity were evaluated after dilution (part 1), cooling (part 2) and freezing and thawing (part 3). The results of the first part of the experiment showed that there were no significant difference between treatments in the motility, progressive motility, viability, membranes and acrosome integrity of spermatozoa, but the results with 2.0 mM BHT were slightly better than obtained with other levels of BHT and control extender. Significantly better results (p<0.05) were observed in the second part of the experiment for cooled spermatozoa characteristics, when extender was supplemented with 2.0 and 3.0 mM BHT. Furthermore, the results obtained in the third part of the experiment indicated that, after freezing and thawing, all evaluated semen characteristics were improved significantly (p<0.05) by increasing BHT levels, with the best results obtained for extender containing 2 mM BHT. Comparison of these results with those of control diluent, the effects of supplementation were significantly (p<0.01) better. However, the higher concentration of BHT (3.0 mM) reduced the motility, acrosomal integrity, viability and hypo-osmotic swelling response of spermatozoa compared to extender containing 2.0 mM BHT. In conclusion, the results obtained in this study showed that the semen quality of rams was improved when BHT was added to extender used before the freezing process.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.2
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• pp.201-209
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• 2000

Objective: This study has been carried out to evaluation the effect of fertilization promoting peptide (FPP) on the kinematic parameters, capacitation and acrosome reaction of the frozen-thawed human spermatozoa. Methods: After FPP treatment, we examined kinematic parameters, capacitation and acrosome reaction, using the methods of computer-aided sperm analysis (CASA) and chlortetracycline (CTC) fluorescence analysis. Results: We have obtained the evidence that FPP can promote the capacitation and inhibit the spontaneous acrosome reaction of frozen-thawed human spermatozoa in vitro. FPP ($25{\sim}100$ nM) induced a significant increase in the proportion of B-pattem capacitated spermatozoa, and a significant decrease in the proportion of F-pattem uncapacitated ones without significant stimulation of acrosomal exocytosis. In the kinematic parameters treatment, FPP treated groups maint3ined higher LIN, BCF and STR than those of control. The VAP, VSL, VCL and ALH were not different. Therefore it is suggested that FPP in human seminal plasma may play a positive role in promoting human sperm function.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.3
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• pp.253-259
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• 1994

A clinical investigation was undertaken on primary male infertility patients of recent 5 years. The results obtained were as follow: 1. Suspective etiologic factors were: 1) testicular failure, 36.1 %; 2) varicocele, 18.7%; 3) endocrine abnormality, 13.5%; 4) obstruction, 13.5%; 5) idiopathic, 10.9%; 6) cryptorchidism, 2.6%; 7) necrospermia, 0.9%. 2. On semen analyses, azoospermia was found in 55.8%, single abnormal parameter in 21.5 %, and multiple/all abnormal parameter in 22.7% of the 163 cases. 3. For the evaluation of the sensitivity and specificity of noninvasive variables in predict in obstruction as the cause of azoospermia in patient who had undergone testicular biopsy, the testicular size and serum follicle-stimulating hormone(FSH) level revealed 100% of sensitivity. 4. Among the 43 patients with a testicular biopsy confirmed diagnosis there was a significant difference in testicular size, ejaculate volume(p<0.0001) and serum FSH(p<0.0001) between patients with testicular failure and those with ductal obstruction. 5. Of 93 treated patients with primary male infertility, 42 were managed by medical treatment including endocrine treament, retrograde ejaculation treatment, infection treatment and observation; 29 were managed by surgical treatment including varicocelectomy, vasovasostomy, vasoepididymostomy and TUR of ejaculatory duct; 20 were managed by sperm preparation treatment including artificial insemination(AI), electroejaculation plus AI and vibration ejaculation plus AI ; 2 were managed by microscopic epididymal sperm aspiration plus IVF, repectively. 6. 42 patients who could be followed-up, 21 patients(50%) impregnated their wives.

• Animal Bioscience
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• v.34 no.5
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• pp.844-854
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• 2021

Objective: The potential of extra virgin olive oil (EVOO), betaine (BET), and ginger (GIN), as natural antioxidants, in reducing negative effects of heat stress on physiological responses, antioxidant capacity, semen quality and fertility of bucks under heat stress were investigated. Methods: Forty adult Animal Production Research Institute line rabbit bucks were distributed randomly into four experimental treatments of ten rabbits each. The first treatment was fed the commercial pellet diet (CPD) without supplementation and served as a control. The other three treatments were fed CPD supplemented with EVOO (300 mg), BET (1,000 mg), and GIN (200 mg) per kg diet for 3 consecutive months during the summer season. Results: Supplementation of EVOO, BET, or GIN improved (p<0.05) the sexual desire, progressive motility, vitality, intact acrosome and membrane integrity, sperm cell concentration, sperm outputs and fertility. Seminal plasma total proteins, globulin, total antioxidant capacity, glutathione and glutathione S-transferase, and initial fructose increased (p<0.05), while total lipids, aspartate and alanine aminotransferases and malondialdehyde decreased (p<0.05) compared with the control. In comparing the natural antioxidants treatments, GIN evoked the largest improvement. Conclusion: The inclusion of GIN (200 mg/kg diet) appeared to improve the sexual desire, semen quality and oxidative stress of bucks. This may be a beneficial supplement for the management of rabbit bucks used in natural mating or artificial insemination.

• Reproductive and Developmental Biology
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• v.35 no.1
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• pp.55-60
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• 2011

The objective of this study was carried out to evaluate the efficiency of sperm collection methods on the post-thaw viability of cat semen. The cat semen was collected by artificial virginal (AV) and electronic ejaculate (EE) methods. The composition of semen extender was consisted of Tris-buffer supplemented with 20% egg yolk and 1% P/S antibiotics in Ext I, and more added 8% glycerol, 1.0% Equex STM paste of total volume in Ext II. The collected semen was adjusted the concentration and then diluted in Ext I for optimal concentration. The diluted semen was cooling to $5^{\circ}C$ temperature in refrigerator for at least 2 hrs and then diluted stepwise with Ext II for at least 1 hrs. After an equilibration for 1 hrs, the cooled semen was packaged in 0.5 ml straw and then freezing on the $LN_2$ vapor over 5 cm above from $LN_2$ and then immersed directly in $LN_2$ for cryopreservation. The frozen semen was thawed in $38^{\circ}C$ water for 15 sec and then evaluated the motility, viability, and morphology. Post-thaw semen were calculated the motility by SMI (sperm motility index). The live-dead sperm was evaluated by Eosin-B and morphological evaluation was by Diff-quik kit staining. The post-thaw concentration ($89{\times}10^6$ /ml vs. $128{\times}10^6$ /ml), viability ($22.6{\pm}10.6%$ vs. $37.1{\pm}26.1%$), morphological normality ($27.0{\pm}50.2%$ vs. $45.6{\pm}123.0%$) of EE and AV groups were not significant different, but the post-thaw motility was significant lower in EE than that in AV group ($53.1{\pm}3.6$ vs. $73.6{\pm}5.7$) (p<0.05). In conclusion, semen collection methods did not significant different between EE and AV groups except of post-thaw motility and so both semen collection methods could be applied in feline semen collection methods.

• Clinical and Experimental Reproductive Medicine
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• v.15 no.1
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• pp.1-9
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• 1988

The contraception practice rate in Korea has steadily increased from 9% in 1964 to 72% in 1987. Accordingly, natural increase rate of population declined from 28.4 per 1,000 in 1964 to 12.4 per 1,000 in 1987. Notably in recent years, the acceptance of surgical contraception services has sharply increased in this country. Now the author analyzes various factors influencing successful vasectomy and vasovasostomy based on his experience in performing more than 8,000 vasectomies and 1,000 vasovasostomies during the past 25 years. These various factors consist of preoperative screening interview, surgical vasectomy technique, vas irrigation technique, open-ended vasectomy, complications from vasectomy, sperm granuloma formation, spermatogenesis and steroidogenesis after vasectomy, immunological consequences after vasectomy, prostatic diseases and vasectomy, late post-vasectomy syndrome, psychological effects after vasectomy, and success rates of vasovasostomy. Besides, the author makes more comments on the results of the clinical trials on new approaches to reversible vas occlusion technique with a new device of Intravasal Thread and to reversible suppression of spermatogenesis with long-acting sex hormones.

• Clinical and Experimental Reproductive Medicine
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• v.19 no.1
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• pp.65-69
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• 1992

The sucess of intrauterine inseminations with washed human spermatozoa was evaluated in 92 patients. In according to indication of insemination, intrauterine inseminations of women with hostile cervical mucus yield a 35.3% and 12.8% with male factors and 27.3% with unexplained infertility. In ovulation induction group with variable agents including clomiphen, human menopausal gonadotropin (HMG) and GnRH analog, the pregnancy rate was 27.8% (22/79) and in natural cycle group, 15.4% (2/13). The fetal loss rate in insemination group was 12.5% (3/24). Multiple pregnancies were 7 cases and ovulation induction were performed in 6 cases among them. Intrauterine insemination with washed human spermatozoa therefore represents an effective and safe procedure selected infertile couples.